EP3726995A1 - Procédé d'identification et d'isolement de composés bioactifs à partir d'extraits d'algues - Google Patents

Procédé d'identification et d'isolement de composés bioactifs à partir d'extraits d'algues

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Publication number
EP3726995A1
EP3726995A1 EP18815767.1A EP18815767A EP3726995A1 EP 3726995 A1 EP3726995 A1 EP 3726995A1 EP 18815767 A EP18815767 A EP 18815767A EP 3726995 A1 EP3726995 A1 EP 3726995A1
Authority
EP
European Patent Office
Prior art keywords
extract
kda
bioactive molecules
filtrate
molecular weight
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP18815767.1A
Other languages
German (de)
English (en)
Inventor
Céline CONAN
Philippe Potin
Anne GUIBOILEAU
Samantha Besse
Jean-Marie Joubert
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Laboratoires Goemar SA
Original Assignee
Laboratoires Goemar SA
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Laboratoires Goemar SA filed Critical Laboratoires Goemar SA
Publication of EP3726995A1 publication Critical patent/EP3726995A1/fr
Pending legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/03—Algae
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P21/00—Plant growth regulators
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/005—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor after treatment of microbial biomass not covered by C12N1/02 - C12N1/08
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5097—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving plant cells

Definitions

  • the present invention relates generally to a method of purifying and isolating bioactive compounds responsible for plant growth stimulation from seaweed extracts.
  • Biostimulants can be used to improve plant nutrition, which impacts yield and quality parameters. Plant biostimulants generally fall within one of these categories i.e. hormone- containing products, plant extract based products, micronutrients based products, amino acid- containing products and humic acid-containing products but may not be strictly restricted to these categories alone. Plant biostimulants are used to treat crops in a commercial setting in view of their ability to increase growth rates, increase stress tolerance, increase photosynthetic rate and increase disease tolerance. Plant biostimulants are generally believed to operate by up- regulating or down-regulating key biological pathway genes.
  • plant biostimulants contain substance(s) and/or micro-organisms whose function when applied to plants or the rhizosphere is to stimulate natural processes to enhance/benefit nutrient uptake, nutrient efficiency, tolerance to abiotic stress, and crop quality. Biostimulants have no direct action against pests, and therefore do not fall within the regulatory framework of pesticides.
  • EBIC European Biostimulant Industry Council
  • Biostimulants are available in a variety of formulations and with varying ingredients but are generally classified on the basis of their source and content. These groups include humic substances (HS), and amino acid containing products (AACP).
  • HS humic substances
  • AACP amino acid containing products
  • Biostimulants are available in a variety of formulations and with varying ingredients but are generally classified into seven main groups on the basis of their source and content. These groups include humic substances (humic and fluvic acids), protein hydrolysates and other N-containing compounds, seaweed extracts and botanicals, chitosan and other biopolymers, inorganic compounds, beneficial fungi and beneficial bacteria.
  • humic substances humic and fluvic acids
  • protein hydrolysates and other N-containing compounds include seaweed extracts and botanicals, chitosan and other biopolymers, inorganic compounds, beneficial fungi and beneficial bacteria.
  • seaweed extracts While the physiological effects of seaweed extracts on plant defenses and plant growth has been examined, little is known about the particular bioactive compounds in seaweed extracts that are responsible for these plant stimulants. It would be desirable to accelerate the identification of these bioactive components in algae, including, for example, brown algae extracts.
  • Phaeophyceae or brown algae are a large group of mostly marine multicellular algae, including many types of seaweed located in both Hemisphere waters. They play an important role in marine environments, both as food and as habitats. Many brown algae, such as members of the order Fucales, commonly grow along rocky seashores. Worldwide, over 1,500 species of brown algae are known. Some species, such as Ascophyllum nodosum, are important in commercial use and have environmental impact as well.
  • U.S. Pat. No. 7,611,716 to Michailovna et al describes a method of processing seaweed to obtain, in a single process, extracts comprising acidic and neutral polysaccharides and an extract comprising low molecular weight biologically active compounds that can be used in medicine, food, perfumery and the cosmetic industry.
  • the reference only describes a method of processing seaweed and does not provide any way of identifying potential plant biostimulant compounds contained therein.
  • U.S. Pat. No. 3,856,569 to Strong describes a method of purifying and concentration of the desirable polyssacharide such as carrageenan or alginate from aqueous solutions derived from marine algae (Rhodophyceae and Phaeophyceae) by subjecting the solutions to ultrafiltration.
  • this reference only provides a method of processing seaweed and does not provide any way of identifying biostimulant compounds contained therein.
  • the present invention relates generally to one or more bioactive molecules isolated from an algae species, the one or more bioactive molecules having a molecular weight in the range of about 0.15 kDa to about 1.0 kDa.
  • the present invention relates generally to a method of isolating and purifying bioactive compounds in an extract obtained from seaweed, the method comprising the steps of:
  • Figures la and lb depict Size Exclusion Chromatography (SEC) fractionation performed on filtered RM-3496 extract and a chromatogram of standards injected on the SEC to evaluate the average molecular weights of the molecules presented in the different fractions.
  • Figure 2 depicts boxplots showing the efficacy of SEC fractionation of RM-3496 on lettuces.
  • Figures 3 a and 3b depict boxplots showing the fresh shoot weights and the fresh root weights of in-vitro cultures of Arabidopsis thaliana treated with the F3 fraction as compared with the untreated control, the RM-3496 extract and the rebuilt RM-3496 extract.
  • Figure 4 depicts a view of the ultrafiltration process in accordance with one aspect of the present invention.
  • Figure 5 depicts boxplots showing the fresh shoot weights of lettuces treated with various ultrafiltrated fractions, retentates and extracts.
  • Figure 6 depicts boxplots showing the fresh shoot weights of wheats treated with various ultrafiltrated fractions, retentates and extracts.
  • plant“biostimulant” is an organic material that contains substance(s) and/or micro-organisms whose function when applied to plants or the rhizosphere is to stimulate natural processes to enhance/benefit nutrient uptake, nutrient efficiency, tolerance to abiotic stress, and crop quality.
  • the term“ about” refers to a measurable value such as a parameter, an amount, a temporal duration, and the like and is meant to include variations of +/-l5% or less, preferably variations of +/-l0% or less, more preferably variations of +1-5% or less, even more preferably variations of +/-l% or less, and still more preferably variations of +/-0.l% or less of and from the particularly recited value, in so far as such variations are appropriate to perform in the invention described herein. Furthermore, it is also to be understood that the value to which the modifier“about” refers is itself specifically disclosed herein.
  • the present invention describes a method of purifying and isolating biostimulant compounds from extracts derived from seaweed that are capable of increasing growth rates and yields of a wide range of crops.
  • the present invention provides a method of purifying the bioactive compounds responsible for plant growth stimulation in seaweed extracts by the metabolomics profiling of the seaweed extracts.
  • the present invention relates generally to one or more bioactive molecules isolated from an algal species, the one or more bioactive molecules having a molecular weight in the range of about 0.15 kDa to about 1.0 kDa.
  • the one or more bioactive molecules are ones that are capable of improving plant growth.
  • the algal species is a brown algal species.
  • the brown algae may comprise an algal species selected from the group comprising Ascophyllum nodosum, Fucus vesiculosus, Sargassum sp., and combinations of one or more of the foregoing.
  • the one or more bioactive molecules do not comprise a sulfated polysaccharide or laminarin.
  • the present invention also relates generally to a method of improving plant growth, the method comprising the step of applying a composition comprising the isolated one or more bioactive molecules to at least one of soil, a plant, or a seed.
  • Improving plant growth includes at least one of the following: promoting seed germination, stimulating root development, prolonging a vegetative period, increasing a period of production, or increasing a period of harvest.
  • the present invention also relates generally to a method of isolating and purifying bioactive compounds in an extract obtained from seaweed, the method comprising the steps of:
  • the method further comprises the step of evaluating the bioactivity of the first filtrate fraction and the additional filtrate fractions to determine their efficacy on plant growth.
  • the efficacy on plant growth may include at least one of the following: promoting seed germination, stimulating root development, prolonging a vegetative period, increasing a period of production, or increasing a period of harvest.
  • the extract is produced from a brown algal species.
  • the extract may be obtained from Ascophyllum nodosum, Fucus vesiculosus, or Sargassum sp. algae.
  • the retentate comprises active molecules selected from the group consisting of sulfated polysaccharides and laminarin, which are active molecules capable of alleviating abiotic stress, such as salt excess, in crops.
  • the first filtrate comprises bioactive molecules having a molecular weight in the range of about 0.15 kDa to about 1.0 kDa.
  • the ultrafiltration membrane may have a molecular weight cutoff (MWCO) of less than 3 kDa, preferably a MWCO of less than 2 kDa, and most preferably a MWCO of less than 1 kDa.
  • MWCO molecular weight cutoff
  • Ascophyllum nodosum (rockweed) is a brown algal fucoid species found in the North Atlantic Ocean and has been used as a source of biostimulant for agricultural crops in order to improve plant growth, plant productivity and food quality.
  • SEC Size Exclusion Chromatography
  • GF Gel Filtration Chromatography
  • the seaweed extract (RM-3496) was fractionated by the SEC fractionation process and the molecules were eluted according to their size (or molecular weights) as shown in Figure l.a.
  • ASuperdex30® resin (GE Healthcare, Bjorkgatan, Sweden) was used to ensure a good separation of molecules with a molecular weight below 10 kDa.
  • the molecular weights of the molecules decrease from the first fraction (i.e., Fl) to the last fraction (i.e., F6).
  • the fractions Fl and F2 were constituted by the larger molecules that flow through the column faster than the salts and the very low molecular weight molecules are eluted in fractions F5 and F6.
  • fraction F3 contained molecules with molecular weights smaller than lkDa to about 0.l8kDa
  • Fraction F4 contained molecules with molecular weights smaller than about 0.2kDa such as Mannitol (l82.2Da)
  • Fractions F5 and F6 contained salts and molecules very low molecular weights such as glycine (75Da).
  • the NMR spectra of the different fractions confirmed the presence of sulfated polysaccharides (fucan polymers) in the first fraction Fl while the second fraction F2 contains laminarin (from about 3 to about 4kDa) and the fraction four F4 contains mannitol (182.2 Da).
  • the last two fractions F5 and F6 contained very low molecular weight molecules and salts.
  • the combination of the different techniques used to desalt the RM-2705 extract provided information about the physicochemical properties of the bioactive molecule(s).
  • the bioactive molecules are polar and their molecular weights range from about 0.15 to about 1 kDa.
  • this information excludes, from the growth- promoting bioactive polymers, fucan polymers which are the major sulfated polysaccharides in Ascophyllum nodosum acidic extract, laminarin (from about 3 to about 4kDa), a beta-l,3-glucan elicitor, and mannitol (182 Da), a polyol that can represent up to about 8-10% of the extract by dry weight.
  • the fraction F3 displayed a strong growth-stimulating activity whereas fractions Fl and F2 were inactive. Flowever, during the development of the latter bioassay, it was shown that in the presence of salt (100 mM NaCl), the fraction F3 was no longer active to stimulate growth, whereas Fl and F2 displayed similar effects, and that the whole RM-2705 extract confer salt tolerance.
  • salt 100 mM NaCl
  • Fl and F2 displayed similar effects
  • the whole RM-2705 extract confer salt tolerance.
  • UF ultrafiltration
  • MWCO molecular weight cutoffs
  • a seaweed extract may be ultrafiltrated using a 1 kDa MWCO membrane.
  • the present invention provides a method of purifying a biostimulant composition derived from a seaweed extract comprising a step of ultrafiltration using a semi-permeable ultrafiltration membrane to separate the molecules of interest from the rest of the mixture according to their molecular weight, size and shape.
  • the ultrafiltration step may be carried out using ultrafiltration equipment in which a seaweed extract solution comprising between about 1% by wt. and about 15% by wt. dry matter, more preferably between about 2% by wt. and about 7% by weight dry matter, is subjected to ultrafiltration using a membrane with a suitable molecular weight cutoff (MWCO).
  • MWCO molecular weight cutoff
  • the ultrafiltration process involves tangential ultrafiltration.
  • the filtrate is collected for its biostimulant properties while recirculating the retentate (or concentrate), which is left apart for other applications at the end of the process.
  • a further purification of the retentate (or concentrate) can be achieved by the addition of fresh water at a rate corresponding to that at which water, together with molecules having a molecular weight less than or equal to 1 kDa is removed from the ultra- filtrate.
  • ultrafiltration can be carried by a process in which the solution reservoir (1) is charged with a batch of seaweed extract.
  • the solution is circulated by line (2) and pump (3) into an inlet manifold (4) of an ultrafiltration unit (5).
  • the ultrafiltration unit (5) comprises one or more cartridges arranged in parallel to provide the appropriate ultrafiltration membrane area.
  • the ultra-filtrate then exits the ultrafiltration unit (5) via outlet line (6) and is collected in tank (7).
  • the ultrafiltration concentrate exits the ultrafiltration unit (5) via outlet manifold (8) and is returned via line (9) to the solution reservoir (1).
  • the membrane contained in the ultrafiltration unit (5) may be polymeric or ceramic type membrane.
  • the membrane comprises a tubular ceramic membrane comprising a plurality of channels.
  • the membrane may contain between about 15 and about 50 channels, more preferably between about 19 and about 39 channels and may have a length between about 50 and about 150 cm.
  • spiral membranes and crossflow membranes may also be used in the practice of the invention.
  • the membrane area is generally between about 0.20 and about 0.6 m , more preferably between about 0.30 and 0.40 m 2 .
  • the retentate is rinsed several times to remove the major portion of molecules with a molecular weight smaller than the cutoff of the membrane.
  • the ultra-filtrates contain molecules with a molecular weight smaller than that of the cutoff membrane. In one instance the cutoff is 3 kDa, more preferably 2 kDa, and still most preferably 1 kDa.
  • the molecules that are contained in the ultra-filtrates display low molecular weights smaller than the MWCO, e.g., 1 kDa, and are commonly referred to as metabolites.
  • the retentates contain molecules with molecular weights larger than the cutoff membranes, e.g., 1 kDa.
  • the molecules that are contained in the retentate display high molecular weights (e.g., Laminarin from about 3 to about 4 kDa or Fucoidans higher than about 30 kDa and other brown algal high molecular weight biopolymers).
  • high molecular weights e.g., Laminarin from about 3 to about 4 kDa or Fucoidans higher than about 30 kDa and other brown algal high molecular weight biopolymers.
  • algal species from the order of Fucales have been found to display a promising activity and can be subjected to the methods described herein.
  • algal species include, but are not limited to, species of the families of Fucaceae, Sargassaceae and Durveilleaceae.
  • Fucales and Laminariales orders include, but are not limited to Ascoseirales, Asterocladales, Desmarestiales, Dictyotales, Dictyotophycidae, Discosporangiales, Discosporangiophycidae, Ectocarpales, Fucales, Fucophycidae, Ishigeales, Ishigeophycidae, Faminariales, Nemodermatales, Onslowiales, Phaeophyceae ordo incertae sedis, Phaeosiphoniellales, Ralfsiales, Scytothamnales, Sphacelariales, Sporochnales, Stschapoviales, Syringodermatales, Tilopteridales, among others.
  • the present invention is described and shown to demonstrate positive results on algal species from the order Fucales, the method is not limited to these algal species and can also be used to isolate and analyze filtrates of any algae or other species that may act as biostimulants to determine bioactivity of such filtrates.
  • the term“filtrate” refers to filtrate and ultra-filtrates obtained after one or more ultrafiltrations through the ultrafiltration unit.
  • retentate refers to retentate without flushing and retentates obtained after one or more flushings.
  • a RM-3496 extract was ultrafiltrated at the laboratory scale with a 1 kDa MWCO membrane and the retentate was rinsed three times with water.
  • the ultra-filtrate, containing molecules with molecular weights smaller than 1 kDa, and the retentate, containing molecules with molecular weights larger than 1 kDa were tested on lettuces and wheat.
  • the GF142 and GS142 extracts (available from Faboratoires Goemar) were manufactured with the same process from Fucus vesiculosus and Sargassum natans respectively.
  • RM-3496 manufactured by Laboratoires Goemar from Ascophyllum nodosum extract and two other seaweed extracts (GF142 and GS142, manufactured by Laboratoires Goemar from Fucus vesiculosus and Sargasssum natans respectively) were subjected to ultrafiltration and evaluated for their biostimulant properties.
  • the total dry weights of the filtrates was about 80% of the RM-3496 extract and the retentate was about 20% of the RM-3496 extract.
  • the lettuce growth experiments were performed with seeds of lettuces Lactuca saliva ecotypes Fabietto or Janero (available from Voltz, Colmar, France). Lettuces were grown in a growth-chamber, on a rotary table to obtain plant phenotype as homogeneous as possible for any condition of treatment. Plants were grown under high pressure iodide-sodium lamps with a photosynthetically active radiation of 150 ⁇ 10 pmol of photons. m-2.s-l, a thermo-period of 20/18°C (day/night) and a long-day photoperiod of l6h light.
  • Seeds were first surface-sterilized and were sown in squared Petri dishes containing Half-strength Murashige and Skoog (MS) basal medium supplemented with 1% (w/v) of sucrose (30mM) and 0.6% (w/v) of PhytagelTM.
  • Petri dishes were grown under a cool fluorescent light with an intensity of 225 ⁇ 10 pmol photons.m-2.s-l, with a long-day photoperiod of l6h light at 2l°C ⁇ 0.5 °C.
  • the location of Petri plates under the neon lamps were changed every day and this all along experiment to randomize the experiment.
  • the wheat growth experiments were performed with seeds of winter wheat ( Triticum aestivum L.) variety Altigo (available from Limagrain, Saint-Beauzire, France). Wheats were grown in a growth chamber on a rotary table to obtain for each condition plant phenotype as homogenous as possible. In order to control the nutrient inputs to the plants, seeds of wheat were grown in vermiculite pots. The plants were grown in the growth-chamber under high pressure iodide-sodium lamps with a photosynthetically active radiation of 150+/- 10 pmol of photons. m .s and a thermo-period of 22/l8°C with a long day photoperiod of 16 hours. Ten days after sowing, homogeneous plants were distributed in different trays; 6 plants per tray and two trays per condition. The plants were watered three times per week with the same commercial nutritive solution used for lettuce experiments.
  • the efficacy of the different fractions and extracts on plant growth stimulation was evaluated by a statistical approach. Indeed, for each bioassay shown, the normality of the data was first checked with Shapiro-Wilk normality tests, with the Q-Q plots and with the histograms of density. The Homoscedasticity of these data was also checked with the Barlett’s test, prior to performed parametric tests on these data. Several bioassays (three to four independent repetitions in time) were carried out to assess the different treatments on plant growth stimulation with a number N of plants by Treatment.
  • a parametric two-way analysis of variance (two-way Anova) was then performed on the data to determine if there was a significant difference (with an alpha error of 5%) between the means of the different treatments for each bioassay and between the means of each treatment for the different bioassays carried out.
  • a parametric post-hoc HSD Tukey’s test or multiple pairwise comparison was performed on the data to range and define what means were significantly different from each other. Tukey’s test results are shown on the boxplots with bold letters. The means of treatments which are significantly different from each other display different bold letters. These means are depicted on each boxplot by a dot.
  • the compounds described herein can be used on various crops including, for example soybeans, corn, cereals (i.e., wheat, barley, rye, and oats), rapeseed, canola, sunflower, sugar beet, potatoes, dry pulses (i.e., lentils, dry beans, etc.), sugarcane, fruiting vegetables, including tomatoes, eggplant, peppers, cucurbits, etc., bulb vegetables, including onions and leeks, head and leafy vegetables, including lettuce, spinach and celery, brassicas, stone fruits, pome fruits, citrus, coffee, cocoa, nut trees, berries, grapes (tables and vines), among others.
  • soybeans i.e., wheat, barley, rye, and oats
  • rapeseed canola
  • sunflower sugar beet
  • sugar beet i.e., lentils, dry beans, etc.
  • sugarcane i.e., lentils, dry beans, etc.
  • sugarcane i.e

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Abstract

L'invention concerne un procédé d'isolement et de purification de composés bioactifs dans un extrait obtenu à partir d'algues. Le procédé comprend les étapes consistant à : (a) faire circuler l'extrait à travers une membrane d'ultrafiltration ayant un seuil de poids moléculaire approprié ; (b) collecter le filtrat de l'extrait pour obtenir une première fraction de filtrat et un rétentat ; et (c) rincer le rétentat pour obtenir une ou plusieurs fractions de filtrat supplémentaires. La bioactivité de la première fraction de filtrat et des fractions de filtrat supplémentaires peut ensuite être évaluée pour déterminer leur efficacité sur la croissance des plantes. L'invention concerne également une ou plusieurs molécules bioactives isolées d'une espèce algale, la ou les molécules bioactives ayant un poids moléculaire dans la plage d'environ 0,15 k Da à environ 1,0 k Da et étant capables de renforcer ou d'améliorer la croissance des plantes.
EP18815767.1A 2017-12-18 2018-12-17 Procédé d'identification et d'isolement de composés bioactifs à partir d'extraits d'algues Pending EP3726995A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FR1762345A FR3074998B1 (fr) 2017-12-18 2017-12-18 Procede pour identifier et isoler des composes bioactifs a partir d'extraits d'algues
PCT/EP2018/085254 WO2019121539A1 (fr) 2017-12-18 2018-12-17 Procédé d'identification et d'isolement de composés bioactifs à partir d'extraits d'algues

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EP3726995A1 true EP3726995A1 (fr) 2020-10-28

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US (1) US20200329714A1 (fr)
EP (1) EP3726995A1 (fr)
JP (1) JP7335245B2 (fr)
KR (2) KR20230003625A (fr)
CN (1) CN111526721A (fr)
AR (1) AR113960A1 (fr)
AU (1) AU2018391577B2 (fr)
BR (1) BR112020012394A2 (fr)
CA (1) CA3085673A1 (fr)
CL (1) CL2020001617A1 (fr)
CO (1) CO2020007276A2 (fr)
CR (1) CR20200318A (fr)
EA (1) EA202091492A8 (fr)
FR (1) FR3074998B1 (fr)
NI (1) NI202000046A (fr)
PH (1) PH12020550866A1 (fr)
UA (1) UA124910C2 (fr)
WO (1) WO2019121539A1 (fr)
ZA (1) ZA202003638B (fr)

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WO2021258146A1 (fr) * 2020-06-24 2021-12-30 University Of The Sunshine Coast Composés bioactifs issus d'algues et procédé

Family Cites Families (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA980261A (en) 1972-08-16 1975-12-23 Uniroyal Ltd. Process for the purification and concentration of solutions derived from marine algae
US4342751A (en) * 1981-03-09 1982-08-03 Eli Lilly And Company Majusculamide C
US4608077A (en) * 1985-04-05 1986-08-26 Schwede John G Process for promoting and regulating plant growth with caulerpin
FR2693454B1 (fr) * 1992-07-07 1994-10-07 Goemar Laboratoires Compositions à usage agricole contenant de la laminarine; procédé de culture de plantes utilisant ces compositions.
JP4236016B2 (ja) * 1998-03-12 2009-03-11 ヤマキ株式会社 アゾジリキソフラノシドアデニン誘導体
EP1318825A2 (fr) * 2000-07-26 2003-06-18 Vitaplant AG Extrait de plante
RU2240816C1 (ru) 2003-07-28 2004-11-27 Тихоокеанский институт биоорганической химии Дальневосточного отделения РАН Способ комплексной переработки бурых водорослей с получением препаратов для медицины и косметологии
JP2005170836A (ja) * 2003-12-10 2005-06-30 Riken Vitamin Co Ltd 海藻抽出物およびそれを含むリパーゼ阻害剤
EP1778219A4 (fr) * 2004-08-17 2010-06-30 Ocean Nutrition Canada Ltd Compositions d'ascophyllum et procedes
RU2337571C2 (ru) * 2006-08-11 2008-11-10 Мурманский морской биологический институт Кольского научного центра Российской академии наук (ММБИ КНЦ РАН) Способ комплексной переработки фукусовых водорослей (варианты)
JP5254572B2 (ja) * 2007-06-22 2013-08-07 エステー株式会社 精製カラギーナン及びその製造方法
CN101580555B (zh) * 2009-05-18 2012-02-29 北京雷力联合海洋生物科技有限公司 一种不同分子量范围的岩藻多糖的制备方法
CN102228125B (zh) * 2011-05-27 2013-04-03 湖州佳美生物化学制品有限公司 海藻活性肽的制备方法
NL2009482C2 (en) * 2012-09-18 2014-03-19 Stichting Energie Process for mannitol extraction from seaweed.
US20150274791A1 (en) * 2012-10-29 2015-10-01 Matis Ohf. Use of natural antioxidants during enzymatic hydrolysis of aquatic protein to obtain high quality aquatic protein hydrolysates
JP6145858B2 (ja) * 2012-11-02 2017-06-14 伊那食品工業株式会社 植物生育促進剤の製造方法及び植物生育促進方法
RO128888A0 (ro) * 2012-11-27 2013-10-30 Soctech S.A. Compoziţie pentru tratamentul culturilor agricole şi procedeu de obţinere
CN103704652B (zh) * 2013-12-17 2015-08-19 山东洁晶集团股份有限公司 一种海藻调味料的制备方法
CN107212422A (zh) * 2017-06-23 2017-09-29 湖州知维技术服务有限公司 一种海藻活性肽的制备方法

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ZA202003638B (en) 2023-10-25
KR20230003625A (ko) 2023-01-06
NI202000046A (es) 2020-11-16
WO2019121539A1 (fr) 2019-06-27
PH12020550866A1 (en) 2021-05-17
FR3074998B1 (fr) 2020-03-27
AU2018391577B2 (en) 2024-05-09
UA124910C2 (uk) 2021-12-08
EA202091492A8 (ru) 2020-11-30
JP7335245B2 (ja) 2023-08-29
AR113960A1 (es) 2020-07-01
CL2020001617A1 (es) 2021-01-04
CN111526721A (zh) 2020-08-11
EA202091492A1 (ru) 2020-09-09
AU2018391577A1 (en) 2020-06-25
CR20200318A (es) 2020-10-05
CA3085673A1 (fr) 2020-06-12
US20200329714A1 (en) 2020-10-22
BR112020012394A2 (pt) 2020-11-24

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